| The rice blast disease,caused by the filamentous ascomycetous fungus Magnaporthe oryzae,is one of important fungal diseases of rice throughout the world. Recently, control measures like the use of resistant cultivars and application of fungicides have not allowed complete eradication of the disease. Important prerequisite for control of the disease is to know molecular regulation of growth and disease mechanisms. However, the availability of the genome sequences of the fungal pathogen and improved molecular genetic techniques have not only accelerated the discovery of pathogenesis related genes but also help research to find novel targets of chemical control and provide new sight and good idea for designing comprehensive management of Magnaporthe oryzae.Endocytosis is a complex and fundamental transport process, including grid protein, regulatory proteins and various lipid swallowed, the kinase phosphorylation and a large number of process, the reconstruction of the actin cytoskeleton and etc. In yeast cells, the adapter protein Edelp and clathrin first gathered within the plasma membrane swallowing function, followed by Ste2p internalization, extracellular clathrin and a homolog of WASP protein, Las17p began to gather and to promote a Arp2/3actin combination, actin packages promote cell swallowed around the vesicle formation. At this point, Panlp-End3p-Slalp complex in cell swallowed point accumulation, further promote the formation of the hollow vesicle. This process regulated by many kinases, Ark/Prk kinase is one of the important regulating kinase family in this process. As in the rice blast fungus, SNAREs such as MoSec22and MoVam7deletion mutants varying degree influence the process of endocytosis.In order to further understand the action of bacteria cell endocytosis in molecular mechanism and the interaction relations, the pathway of endocytosis related genes in rice blast fungus growth and systemic research on the functions in the pathogenic process. Through screening proteins interacted with MoARKl, a endocytosis gene MoPAN1was found in this article. Through co-IP,we further establish the relationship between both interactions. Adopt the strategy of gene silencing, biological character of the mutants analysis found that MoPANl silence mutants involved in regulating the sensitivity to the different stress factors, spore production quantity reduced, and the spore morphology anomalies, turgor pressure lossing, affected melanin formation, leading to unsuccessful invasion of the host, pathogenic significantly reduced. Meanwhile MoPAN1involved in sexual reproduction and the hydrophobicity of hypha, regulating the extracellular laccase and the activity of peroxidase.Most importantly MoPAN1involved in various physiological formings of cell wall integrity and rice blast fungus cell function regulation, formation of endocytosis and nuclear signal output.Moreover, localization of MoPanl at the hyphae tip was similar to actin protein, and positioning was regulated by both MoSec22and MoVam7of SNARE protein family.Meanwhile, we characterized another endocytosis gene MoEDEl, a homolog of Ede1p in S.cerevisiae, which appears to be encoded by a single copy gene. Like MoPANl, MoEDE1possesses a potential EH (Eps15Homology) domain. Disruption of MoEDEl resulted in low sensitivity to different stress factors, participated in the regulation of asexual spore and mycelium vegetative growth, affected the integrity of its cell wall, and lost in endocytosis, but had no significant effection in pathogenic.MoEdel has the same positioning at top of the hyphae similar to the actin protein. Above results showed that the endocytosis genes has an important regulatory role in the growth,cell wall integrity and polar growth of mycelium of Magnaporthe oryzae, but different endocytic genes differ in terms of function and regulatory mechanism. |
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